间断气体压力对体外培养成人骨髓基质细胞增殖分化的影响  

Influence of discontinuous pressure on the proliferation and differentiation of in vitro cultured adult bone marrow stromal cells

作　　者：胡小毅[1] 罗莉[2] 

机构地区：[1]广西医科大学附属口腔医院综合门诊,广西壮族自治区南宁市530021 [2]广西医科大学附属口腔医院颌面外科,广西壮族自治区南宁市530021

出　　处：《中国临床康复》2005年第46期20-21,T0001,共3页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:取成人的骨髓液分离纯化得骨髓基质细胞进行体外培养,并对细胞施加间断气体压力,观察细胞在加压后的增殖及分化情况。方法:实验于2001-11/2004-04在广西医科大学实验中心完成。取无骨髓疾病患者骨髓经分离纯化后得骨髓基质细胞,接种于孔板内,在原代培养72h时将培养板以封口膜封闭后置于压力装置内,每天施加压力(0.098MPa)8h,按照加压15min→放压15min→加压15min顺序连续9d。另设对照组不加压力,其余培养条件与加压组相同。分别在加压的第3,6,9天进行细胞计数和碱性磷酸酶含量检测。结果:①施加间断压力的骨髓基质细胞和对照组相比其增殖、分化速度快,经统计学分析该差别有统计学意义[第3天:(12.30±1.16)～(12.90±1.10)个/视野,(10.30±1.06)～(11.00±1.05)个/视野;第6天:(14.80±1.75)～(15.90±0.99)个/视野,(12.30±0.95)～(13.40±1.26)个/视野;第9天:(16.20±1.48)～(17.20±1.62)个/视野,(12.80±0.79)～(14.10±0.99)个/视野,P<0.05]。随着加压时间延长,两组增殖、分化速度无明显差别。②培养至第3,6,9天,实验组较对照组具有较强的碱性磷酸酶活性(A)[第3天:0.589±0.084～0.722±0.076,0.511±0.116～0.655±0.072;第6天:0.781±0.107～0.829±0.054,0.648±0.123～0.747±0.077;第9天:0.870±0.096～0.902±0.132,0.645±0.107～0.812±0.188,P<0.05]。结论:合理的体外培养和适当的生物外力可以促进离体培养的成人骨髓基质细胞的增殖和分化。AIM： To in vitro culture the bone marrow stromal cells （MSCs） isolated and purified from adults＇ bone marrow, treat the MSCs with discontinuous pressure, and observe the proliferation and differentiation of the MSCs after compressure. METHODS： The experiment was carried out in the experimental center of Guangxi Medical University between November 2001. The MSCs were isolated and purified from healthy adults＇ bone marrow, and then inoculated in 96-well plates, then the plates were sealed at 72 hours, and discontinuous pressure of 0.098 MPa was given for 8 hours every day following the order of compression for 15 minutes and decompression for 15 minutes and then compression for 15 minutes for 9 continuous days. No pressure was given in the control group, other culture conditions were the same as those in the other groups. Cell counting and content of alkaline phosphatase were detected on the 3^rd, 6^th and 9^th days of pressure. RESULTS： The proliferation and differentiation in the discontinuous pressure groups were faster than those in the control group [3^rd day： （12.30 ±1.16）-（12.90±1.10） per visual sight, （10.30±1.06）-（11.00±1.05） per visual sight; 6^th day： （14.80±1.75）-（15.90±0.99） per visual sight, （12.30±0.95） -（13.40±1.26） per visual sight; 9th day： （16.20±1.48）-（17.20±1.62）] per visual sight, （12.80±0.79）-（14.10±0.99） per visual sight, P 〈 0.05]. With the prolongation of pressure, the velocities of proliferation and differentiation had no obvious differences＇between the two groups. ② Till the 3^rd, 6^th and 9^th days of culture, the activities of alkaline phosphatase in the study group were higher than those in the control group [3^rd day： 0.589±0.084 to 0.722 ±0.076, 0.511±0.116 to 0.655±0.072; 6^th day： 0.781 ±0.107 to 0.829 ±0.054, 0.648±0.123 to 0.747±0.077; 9^th day： 0.870±0.096 to 0.902±0.132, 0.645±0.107 to 0.812±0.188, P 〈 0.05] CONCLUSION： Reasonable in vitro cultivation and suitable

关 键 词：细胞培养 压力 细胞分裂 细胞分化 

分 类 号：R394.2[医药卫生—医学遗传学]